Activated Cell Therapy, Incorporated is a recently formed biotechnology company committed to the development and application of cell based therapeutics, including immunotherapeutic approaches to HIV infection. The company has in-house expertise in cellular immunology, molecular genetics, cell processing and large-scale cell expansion. We will be utilizing this expertise in the proposed SPIRAT for purposes of producing and expanding HIV antigen specific cytolytic T cell clones for adoptive immunotherapy. These clones will be generated from HIV-seronegative donors with technology developed by our collaborators at Stanford University (Project II). We will expand and process these cells under good manufacturing practices for purposes of assessing their anti-viral effects, in vivo, in pilot clinical studies as described in Project I. In addition, we will make use of a highly sensitive technique to determine homing and half-life of donor T cells in the peripheral blood and tissues of the recipient. This technique is based on the highly polymorphic VNTR markers which will allow us to distinguish donor and recipient cells in a quantitative manner. Finally, where applicable we will transduce the CTL clones with a TAR decoy vector to protect the CTL from HIV infection post-infusion. By carefully defining the specificity, potency, and clonality of our expanded HIV specific CTL clones, we should be able to gain significant insight into their impact on viral load and clinical course.